PROJECT SUMMARY Driving Biomedical Projects Portfolio To drive the development of our molecular microscope pipeline, we have selected Driving Biomedical Projects (DBPs) that serve as the first stage in applying and testing NCDIR technologies. Each DBP contains one or more key specific technological roadblocks that have been generally acknowledged by the field and that must be overcome by the development and optimization of technologies at different stages in our pipeline. Collectively, these DBPs are selected to test all the technology in this pipeline: helping develop sample preparation methods and ways to isolate intact native assemblies; ensuring the development and refinement of methods that determine the composition, stoichiometry, morphology and connectivity of isolated complexes; and projects that challenge the development of methods for modeling static structures of large macromolecular machines as well as dynamic processes involving these machines on long time scales. We have also selected these DBPs to represent different biological problems, key pieces of the ?cellular information pathway? that starts with signalling to a cell, leading to reorganization and activation of DNA in chromatin for transcription into RNA, which is processed, packaged, and exported into the cytoplasm where its translation into protein leads to the generation of new assemblies, structures, and organelles. These DBPs are therefore a key metric for the efficacy of our technologies in both solving key technical roadblocks and addressing key biomedical challenges in the research community we serve. In doing so, we will generate new tools, techniques, and reagents for interactomic studies, train DBP members in the molecular microscope pipeline approaches, begin the dissemination of these approaches to a wider group of researchers both through this training and presentation in meetings and papers, and aid the stabilization of these approaches for a sustainable presence in the community. We also expect application of our TR&D technology in the DBPs to generate new challenges, feeding back to help drive new aspects of our ongoing TR&D development and maturation.